Microsoft Word - 2-AzariHamidian-RTL, Gally proof, Last version.doc Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 9 Original Article Larval Habitat Characteristics of Mosquitoes of the Genus Culex (Diptera: Culicidae) in Guilan Province, Iran S Azari-Hamidian School of Public Health, Guilan University of Medical Sciences, Rasht, Iran (Received 17 Oct 2006; accepted 28 Mar 2007) ABSTRACT An investigation was carried out during April to December 2000 to study mosquito fauna and ecology in Guilan Prov- ince of northern Iran. The mosquito larvae were collected by dipping method and larval habitat characteristics recorded ac- cording to hydro-ecological features. In total, 3937 larvae of the genus Culex from 92 larval breeding sites were collected. Six species of the genus Culex; Cx. mimeticus, Cx. pipiens, Cx. theileri, Cx. tritaeniorhynchus, Cx. hortensis, and Cx. terri- tans were identified in the province and respectively comprised 10.3%, 47.2%, 2.2%, 31%, 6.5%, and 2.8% of the samples. Most of the larvae were collected from the natural habitats (75.6%) such as river edges (6.5%), riverbed pools (28.2%), rain pools (47.8%), stream edges (9.4%), grasslands (1.9%), marshes (2.8%), and hoof-prints (3.4%) and others from artificial habitats (24.4%) including rice fields (32.1%), irrigation channels (7.1%), wells (16.4%), discarded concrete tubes (33.1%), discarded tires (11.0%), and agricultural water-storage pools (0.3%). The ecology of Cx. pipiens and Cx. tritaeniorhynchus, which are the most prevalent species and potentially involved in the transmission of many pathogens to humans and domes- ticated animals, must be extensively studied. Keywords: Culex, Ecology, Iran, Larva INTRODUCTION The genus Culex L. includes 25 subgenera and at least 751 species in the world fauna (Service 1993, Reinert 2001). Certain species of the genus Culex are involved in the transmis- sion of the various arboviral and filarial dis- eases to humans and domesticated animals and/or are important for their biting nuisance in different parts of the world (Cranston et al. 1987, Service 1993). Naficy and Saidi (1970) reported Sindbis virus in Iran as well as West Nile virus in Iran and Guilan Province of north- ern Iran. Siavashi and Massoud (1995) reported human cutaneous dirofilariasis in Guilan Prov- ince. There is no information about the vectors of these diseases in Iran. The possibility of some culicine borne arboviral outbreaks like Japanese encephalitis (JE) and Rift Valley fever in the WHO Eastern Mediterranean Region, including Iran, is noteworthy (WHO 2004). The species Cx. tritaeniorhynchus Giles was men- tioned as the vector of JE in Iran and Iraq (Har- bach 1988). Based on the last classification of the tribe Aedini (Reinert et al. 2004) and the genus Anophe- les Meigen (Harbach 2004), Iranian mosquitoes include nine genera and near to seventy species (Zaim and Cranston 1986, Azari-Hamidian et al. 2002b, Sedaghat and Harbach 2005, Azari- Hamidian et al. 2006, Doosti et al. 2006). While providing exact Iranian mosquito checklist needs more investigations, 24 species of the genus Culex have been recorded so far in Iran (Gut- sevich 1943, Lotfi 1970, 1973, 1976, Danilov 1975, Zaim and Cranston 1986, Harbach 1988). The old records of Cx. apicalis Adams in the Palaearctic Region by different investigators, Corresponding author: S Azari-Hamidian, Tel/Fax: +98 131 3229599, E-mail: azari@gums.ac.ir Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 10 including Iran by Gutsevich (1943), were mis- identifications. This species is very similar to Cx. territans Walker and occurs only in the Nearctic Region (Cranston et al. 1987). Cx. im- pudicus Ficalbi was recorded in Iran only by Lotfi (1970, 1973, 1976) and was not men- tioned in the checklist of Iranian Culex by Zaim and Cranston (1986). Zaim and Cranston (1984) noted that the records of Cx. vishnui Theobald in Iran were misidentifications and the correct name is Cx. pseuodovishnui Colless. Also, Culex vagans Wiedemann and Cx. torrentium Martini are hard differentiated from Cx. pipiens L. in the larval stage (Zaim and Cranston 1986, Har- bach 1988), so the records of these species based on the larval stage (Lotfi 1970, 1976) are doubt- ful. Of course, Danilov (1975) recorded Cx. tor- rentium in Rasht, northern Iran. Cx. univittatus Theobald in southwestern Asia and Iran was considered to refer to Cx. perexiguus Theobald. The former species is an Afrotropical one that occurs only in Yemen Republic in south-west- ern Asia (Harbach 1988). At last, Zaim and Cran- ston (1986) mentioned 19 species of the genus Culex in Iran. Almost half of the Iranian mos- quito species (31 species) and seven genera have been reported in Guilan Province (Danilov 1975, Harbach 1988, Azari-Hamidian et al. 2002b, Sedaghat et al. 2003, Gholizadeh et al. 2004). There are more faunistic data about the ge- nus Culex than other culicine mosquitoes in Iran, but the information on their ecology is not great. Dow (1953) presented some notes on mosquitoes including six species of the genus Culex in different areas of Iran such as larval breeding places and associated species. Go- lestani (1967) studied Cx. pipiens (as form mo- lestus Forskal) in the sewage wells of Tehran. Lotfi (1970, 1973, 1976) briefly noted some ecological aspects of the Culex species that she found in Iran including larval breeding places and water temperature and pH. Yaghoobi-Er- shadi et al. (1986) showed the characteristics of the larval breeding sites and associated species of mosquitoes including eight species of the genus Culex in Minab, Hormozgan Province (southern Iran). Zaim (1987a) found six species of this genus in Kashan, Isfahan Province (cen- tral Iran), and noted some aspects of their adult and larval ecology. Zaim (1987b) reviewed the distribution and larval habitat characteristics of the Iranian Culicinae including 19 species of the genus Culex. Mousakazemi et al. (2000) stressed the fauna and ecology of mosquitoes including Cx. pipiens, Cx. theileri Theobald, and Cx. per- exiguus in Zarrin-Shahr and Mobarakeh area of Isfahan Province. Azari-Hamidian et al. (2002a) studied mosquito fauna in Rasht County of Guilan Province, northern Iran, and presented some breeding places and associated species of Cx. mimeticus Noe, Cx. pipiens, Cx. theileri, and Cx. tritaeniorhynchus. The ecological data about Culex larvae in Iran are mostly based on the little information of faunal records, only Yaghoobi-Ershadi et al. (1986) and Zaim (1987b) exactly studied the larval habitat characteristics of the genus, espe- cially based on the specimens of southern Iran. There is little information about the ecology of this genus in northern Iran. This article is the first that specializes in some aspects of the ecology of Culex larvae such as their larval habitat characteristics, water temperature, and associated species in northern Iran. MATERIALS AND METHODS Study area Guilan Province in the Caspian Sea littoral of northern Iran, between Caspian Sea and Al- borz mountain chains has coastal, plain, foot- hill, and mountainous areas with an area of ap- proximately 14700 square kilometers. The prov- ince has a temperate climate and relatively warm- humid summer, and is located between 36o34´- 38o27´ N latitudes and 48o34´-50o36´ E longi- tudes. With about 1200 mm annual rainfall, Guilan Province has the highest rainfall in Iran. The vast forests of Hyrcania and temperate cli- mate of this province provide ideal conditions for mosquitoes to breed. Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 11 Specimen and data collection Guilan Province includes 16 counties. In each county one fixed and three variable sites (including different topographical areas) were randomly selected and larval collection was carried out for 15-20 minutes in the sites during the spring, summer, and autumn seasons in 2000. Larvae were collected from natural breed- ing sites such as river edges, riverbed pools, rain pools, marshes, grasslands, and tree holes and from artificial breeding sites such as rice fields, irrigation channels, wells, discarded tires, and buckets by means of dipper, pipette, and bucket. Physical and biological characteristics of the larval habitats such as habitat situation (permanent or transient, standing or running), habitat kind (natural or artificial), vegetation situation (with or without vegetation), bottom type, sunlight situation (full or partial sunlight or shaded), water situation (clear or turbid), and water temperature were recorded. Taxonomic note Despite collecting and identifying Anophe- les hyrcanus (Pallas) and An. pseudopictus Grassi in the adult stage in Guilan Province (Dow 1953, Azari-Hamidian et al. 2002b), these species have not been differentiated from each other in the larval stage in the present investiga- tions and mentioned only as An. ’hyrcanus’ in this article. Different species of An. maculipen- nis complex have been recorded in the province according to egg pattern and Polymerase Chain Reaction (PCR) (Dow 1953, Azari-Hamidian et al. 2002b, Sedaghat et al. 2003, Gholizadeh et al. 2004), but they have not been differentiated in the larval stage either and are cited as An. maculipennis Meigen sensu lato. The abbrevia- tions for mosquito genera and subgenera follow Reinert (2001) and Reinert et al. (2004). RESULTS A total of 6656 larvae including 1547 anopheline larvae (23.2%) and 5109 culicine larvae (76.8%) were collected from 127 larval breeding sites on 55 occasions. Among 127 lar- val breeding sites 14 (11.0%) included only the subfamily Anophelinae, 37 (29.2%) included only the subfamily Culicinae, and 76 (59.8%) in- cluded both subfamilies Anophelinae and Cu- licinae. Anopheline larvae were found in 90 breeding sites (70.8%) and culicine larvae in 113 (88.9%). During this study, 3937 larvae of the genus Culex were collected from 92 larval breeding sites (72.4% of the total breeding sites and 81.4% of the culicine breeding sites) on 44 occasions (80.0% of the total). Six species of the genus Culex; Cx. (Culex) mimeticus, Cx. (Cux.) pipiens, Cx. (Cux.) theileri, Cx. (Cux.) tritaeniorhynchus, Cx. (Maillotia) hortensis Fi- calbi, and Cx. (Neoculex) territans were found among the samples. The larvae were collected from many different natural habitats (75.6%) such as river edges (6.5%), riverbed pools (28.2%), rain pools (47.8%), stream edges (9.4%), grasslands (1.9%), marshes (2.8%), and hoof-prints (3.4%) and artificial habitats (24.4%) including rice fields (32.1%), irrigation channels (7.1%), wells (16.4%), discarded concrete tubes (33.1%), dis- carded tires (11.0%), and agricultural water- storage pools (0.3%). Abundance percentages of these species are shown in Table 1. Details of the larval habitat characteristics of the six species are shown in Table 2. The association occasions of the Culex species with other col- lected species are shown in Table 3. The asso- ciation percentages of the species with other mosquito larvae are shown in Table 4. Culex (Culex) mimeticus Noe Cx. mimeticus larvae were collected on 12 occasions (21.8% of the total) from 14 larval breeding sites (11.0% of the total and 12.3% of the culicine breeding sites) in June, July, Au- gust, September, October, and November 2000. Maximum and minimum breeding site water tem- peratures were respectively 28 oC and 12 oC, and the mean temperature was 21.6 oC (for 13 larval breeding sites). Culex (Culex) pipiens L. Cx. pipiens larvae were collected on 28 oc- casions (50.9% of the total) from 40 larval breeding sites (31.4% of the total and 35.3% of Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 12 the culicine breeding sites) in April, May, June, July, August, September, October, and Novem- ber 2000. Maximum and minimum breeding site water temperatures were respectively 32 oC and 12 oC, and the mean temperature was 20.1 oC (for 39 larval breeding sites). Culex (Culex) theileri Theobald Cx. theileri larvae were collected on 7 occa- sions (12.7% of the total) from 10 breeding sites (7.8% of the total and 8.8% of the culicine breeding sites) in May, June, July, and August 2000. Maximum and minimum breeding site water temperatures were respectively 33 oC and 20 oC, and the mean temperature was 25.2 oC (for 10 larval breeding sites). Culex (Culex) tritaeniorhynchus Giles Cx. tritaeniorhynchus larvae were collected on 25 occasions (45.4% of the total) from 44 breeding sites (34.6% of the total and 38.9% of the culicine breeding sites) in June, July, Au- gust, September, October, and November 2000. Maximum and minimum breeding site water temperatures were respectively 33 oC and 12 oC, and the mean temperature was 23.5 oC (for 44 larval breeding sites). Culex (Maillotia) hortensis Ficalbi Cx. hortensis larvae were collected on 7 oc- casions (12.7% of the total) from 15 breeding sites (11.8% of the total and 13.2% of the cu- licine breeding sites) in May, August, and Oc- tober 2000. Maximum and minimum breeding site water temperatures were respectively 31 oC and 12 oC, and the mean temperature was 19.6 oC (for 15 larval breeding sites). Culex (Neoculex) territans Walker Cx. territans larvae were collected on 8 oc- casions (14.5% of the total) from 11 breeding sites (8.6% of the total and 9.7% of the culicine breeding sites) in April, June, July, August, Sep- tember, and October 2000. Maximum and mini- mum breeding site water temperatures were re- spectively 32 oC and 16 oC, and the mean tem- perature was 23.0 oC (for 11 larval breeding sites). Table 1. Composition and abundance of the genus Culex larvae in Guilan Province, Iran, Apr to Dec 2000 Category No. Percentage in family (%) Percentage in subfamily (%) Percentage in genus (%) Cx. mimeticus 405 6.08 7.93 10.29 Cx. pipiens 1858 27.92 36.37 47.19 Cx. theileri 88 1.32 1.72 2.24 Cx. tritaeniorhynchus 1220 18.33 23.88 30.99 Cx. hortensis 256 3.85 5.01 6.50 Cx. territans 110 1.65 2.15 2.79 Total 3937 59.15 77.06 100 Culicinae 5109 76.76 - - Culicidae 6656 - - - Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 13 Table 2. Larval habitat characteristics and occurrence percentages of larvae of the genus Culex in Guilan Province, Iran, Apr to Dec 2000 Larval breeding site- characteristics and habitats Cx. mimeticus (%) Cx. pipiens (%) Cx. theileri (%) Cx. tritaeniorhynchus (%) Cx. hortensis (%) Cx. territans (%) Culex (%) A) Habitat situation 1. Permanent 0.49 19.05 - 0.74 4.3 3.64 9.67 2. Transient 99.51 80.95 100 99.26 95.7 96.36 90.33 3. Slow-running water 0.25 9.31 - 0.49 1.17 2.73 4.73 4. Standing water 99.75 90.69 100 99.51 98.83 97.27 96.27 B) Vegetation situation 1. Without vegetation 7.40 68.14 1.14 24.18 76.95 51.82 46.89 2. With vegetation 92.60 31.86 98.86 75.82 23.05 48.18 53.11 2a) Out of water 14.40 98.64 83.90 72.21 35.59 13.20 67.28 2b) Water surface - - - 1.51 - - 0.66 2c) Beneath of water surface 86.13 35.97 19.54 38.48 66.10 86.80 47.53 C) Bottom type 1. Mud 22.47 82.19 84.09 55.24 59.45 57.8 62.73 2. Sand 15.06 - - 16.57 39.37 15.6 11.32 3. Gravel 62.47 17.81 15.91 28.19 1.18 26.6 25.95 D) Water situation 1. Turbid - 2.83 - 0.25 - - 1.35 2. Clear 100 97.17 100 99.75 100 100 98.65 E) Sunlight situation 1. Full sunlight 99.75 35.67 95.45 84.09 36.33 6.36 57.79 2. Partial sunlight 0.25 12.43 - 3.3 28.12 35.46 9.74 3. Shaded - 51.19 4.55 12.61 35.55 58.18 32.47 F) Habitat kind 1. Natural habitat 88.15 67.30 78.62 80.59 84.77 98.18 75.57 1a. River edge 14.85 0.85 - 8.95 17.98 12.04 6.53 1b. Riverbed pool 83.19 8.29 66.40 26.15 30.41 53.70 28.25 1c. Stream edge 0.28 13.40 - 11.81 - 2.78 9.43 1d. Grassland - - 13.60 4.36 - - 1.85 1e. Marsh - 6.35 - - - - 2.76 1f. Rain pool 1.68 63.36 20 48.62 51.61 31.48 47.78 1g. Hoof-print - 7.75 - 0.11 - - 3.40 2. Artificial (man-made) habitat 11.85 32.70 21.38 19.41 15.23 1.82 24.43 2a. Rice field 100 7.81 100 66.19 94.87 50 32.08 2b. Rice irrigation channel - - - 32.38 - - 7.08 2c. Well - 24.56 - - 5.13 50 16.36 2d. Discarded concrete tube - 50.72 - - - - 33.13 2e. Discarded tire - 16.91 - - - - 11.04 2f. Water-storage pool - - - 1.43 - - 0.31 Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 14 Table 3. The association occasions of Culex larvae with different mosquito larvae in Guilan Province, Iran, Apr to Dec 2000 Table 4. Association percentages of Culex larvae with different mosquito species larvae in Guilan Province, Iran, Apr to Dec 2000 Species T otal occasions A n. claviger A n.’hyrcanus’ A n. m aculipennis s.l. A n. plum beus A n. superpictus A e. vexans C x. m im eticus C x. pipiens C x. theileri C x. tritaeniorhynchus C x. hortensis C x. territans C s. longiareolata C s. m orsitans C s. annulata U r. unguiculata Cx. mimeticus 14 - 12 10 - 2 3 - 4 - 5 1 1 - 1 1 1 Cx. pipiens 40 2 13 14 1 1 4 4 9 3 12 4 5 3 1 3 - Cx. theileri 10 - 6 8 - 2 - - 2 - 8 1 - 1 - - - Cx. tritaeniorhynchus 44 1 31 30 - 5 1 5 12 8 3 4 3 2 - - - Cx. hortensis 15 4 4 2 - 2 1 1 4 1 4 1 - 5 - 2 - Cx. territans 11 4 5 5 - - 2 1 5 - 3 - - - 1 - - Species association Frequency (%) Cx. mimeticus An.’hyrcanus’, An. maculipennis s.l., Cx. tritaeniorhynchus 41.98 An.’hyrcanus’, An. maculipennis s.l. 16.05 An.’hyrcanus’, An. maculipennis s.l., An. superpictus, Cx. tritaeniorhynchus 12.34 An. maculipennis s.l. 11.85 Cx. hortensis, Cx. pipiens 9.38 An.’hyrcanus’, An. maculipennis s.l., Cx. pipiens 7.16 An.’hyrcanus’, An. superpictus, Ur. unguiculata 0.74 An.’hyrcanus’, Cx. pipiens, Cx. territans, Cx. tritaeniorhynchus 0.25 An.’hyrcanus’, Ae. vexans, Cx. pipiens, Cs. annulata, Cs. morsitans 0.25 Total 100 Cx. pipiens Alone 31.16 An. maculipennis s.l. 11.95 An.’hyrcanus’, Cx. hortensis, Cx. tritaeniorhynchus 10.55 Cx. tritaeniorhynchus 6.08 An.’hyrcanus’, An. maculipennis s.l., Cx. theileri, Cx. tritaeniorhynchus 5.49 An.’hyrcanus’, Ae. vexans, Cs. annulata 4.41 An. claviger, An.’hyrcanus’, An. maculipennis s.l., Ae. vexans 3.93 Cx. tritaeniorhynchus, Cs. longiareolata 3.40 An.’hyrcanus’, An. maculipennis s.l., Cx. mimeticus 3.34 An. plumbeus 2.91 An.’hyrcanus’, An. maculipennis s.l., Cx. territans 2.31 Cx. hortensis, Cs. annulata 2.26 An. maculipennis s.l., Cx. tritaeniorhynchus 2.26 Ae. vexans 1.72 An.’hyrcanus’ 1.61 An.’hyrcanus’, Ae. vexans, Cx. mimeticus, Cs. annulata, Cs. morsitans 1.45 An. claviger, Cx. territans 0.97 An.’hyrcanus’, An. maculipennis s.l., Cx. tritaeniorhynchus 0.97 An. maculipennis s.l., Cx. territans 0.86 An.’hyrcanus’, An. maculipennis s.l., Cx. theileri, Cx. tritaeniorhynchus, Cs. longiareolata 0.7 Cx. territans 0.59 An.’hyrcanus’, Cx. mimeticus, Cx. territans, Cx. tritaeniorhynchus 0.38 Cs. longiareolata 0.32 An.’hyrcanus’, An. maculipennis s.l., Cx. theileri 0.16 An.’hyrcanus’, An. superpictus, Cx. hortensis, Cx. tritaeniorhynchus 0.11 Cx. mimeticus, Cx. hortensis 0.11 Total 100 Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 15 Table 4. Continue… Cx. theileri An.’hyrcanus’, An. maculipennis s.l., Cx. tritaeniorhynchus 37.50 An.’hyrcanus’, An. maculipennis s.l., An. superpictus, Cx. tritaeniorhynchus 19.32 An.’hyrcanus’, Cx. hortensis, Cx. tritaeniorhynchus 18.18 An. maculipennis s.l., Cx. tritaeniorhynchus 11.36 An.’hyrcanus’, An. maculipennis s.l., Cx. pipiens 3.41 An. superpictus, Cx. tritaeniorhynchus 3.41 An.’hyrcanus’, An. maculipennis s.l., Cx. pipiens, Cx. tritaeniorhynchus 3.41 An. maculipennis s.l. 2.27 An.’hyrcanus’, An. maculipennis s.l., Cx. pipiens, Cx. tritaeniorhynchus, Cs. longiareolata 1.14 Total 100 Cx. tritaeniorhynchus An.’hyrcanus’, An. maculipennis s.l. 30.90 An.’hyrcanus’ 11.97 Cx. pipiens 9.51 Alone 6.64 An.’hyrcanus’, An. maculipennis s.l., Cx. theileri 5.90 An.’hyrcanus’, Cx. hortensis, Cx. theileri 5.24 An.’hyrcanus’, An. maculipennis s.l., Cx. pipiens, Cx. theileri, Cs. longiareolata 4.92 An.’hyrcanus’, An. maculipennis s.l., An. superpictus, Cx. mimeticus 3.61 An.’hyrcanus’, An. maculipennis s.l., Cx. mimeticus 3.36 An.’hyrcanus’, An. maculipennis s.l., An. superpictus, Cx. theileri 3.11 An. maculipennis s.l., An. superpictus 2.95 An.’hyrcanus’, An. superpictus, Cx. hortensis, Cx. pipiens 2.30 An. maculipennis s.l., Cx. pipiens 2.13 An. superpictus, Cx. theileri 1.89 An.’hyrcanus’, Ae. vexans 1.56 An.’hyrcanus’, An. maculipennis s.l., Cx. hortensis 1.15 An.’hyrcanus’, An. maculipennis s.l., Cx. territans 0.82 An.’hyrcanus’, An. maculipennis s.l., Cx. pipiens, Cx. theileri 0.74 An.’hyrcanus’, Cx. mimeticus, Cx. pipiens, Cx. territans 0.49 An.’hyrcanus’, An. maculipennis s.l., Cx. pipiens 0.41 An. claviger, An. maculipennis s.l., Cx. territans 0.16 An.’hyrcanus’, Cx. hortensis, Cx. pipiens 0.16 Cx. pipiens, Cs. longiareolata 0.08 Total 100 Cx. hortensis Cs. longiareolata 30.47 An. claviger 14.85 Ae. vexans 14.46 An. maculipennis s.l. 14.06 An. claviger, Cs. longiareolata, Cs. annulata 7.81 An. claviger, Cs. longiareolata 6.25 An. claviger, An. superpictus, Cs. longiareolata 3.52 An.’hyrcanus’, Cx. pipiens, Cx. tritaeniorhynchus 2.34 An.’hyrcanus’, Cx. theileri, Cx. tritaeniorhynchus 1.56 An.’hyrcanus’, An. superpictus, Cx. pipiens, Cx. tritaeniorhynchus 1.56 Alone 1.17 Cx. pipiens, Cs. annulata 0.78 Cx. mimeticus, Cx. pipiens 0.78 An.’hyrcanus’, An. maculipennis s.l., Cx. tritaeniorhynchus 0.39 Total 100 Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 16 Table 4. Continue… DISCUSSION Results of this study revealed that the genus Culex species laid their eggs in many different natural and artificial habitats. In parallel to the present research, Service (1993) stressed this genus is, by far, the largest, most common, and most important genus of the tribe Culicini. In these investigations, the species of this genus were the most abundant species of mosquitoes in the studied area (Table 1). Cx. mimeticus larvae are found in pooled streams, springs, and pools (Horsfall 1955). Dow (1953) collected the larvae in shallow pebbly pools in northern Iran. Lotfi (1970, 1976) found it in river edges and riverbed pools with water temperature 18-24 oC and pH 6-7.5. Some other breeding places of this species are pools in swampy ground, irrigation ditches, and animal hoof-prints (Harbach 1988). Zaim (1987b) found this species mostly in natural habitats (88.7%), and in man-made ones such as over- flow water and rice fields. Azari-Hamidian et al. (2002a) observed it only in natural habitats. This phenomenon was nearly seen in this study either, and a few Cx. mimeticus larvae (11.8%) found only in rice fields as artificial habitat (Table 2). Harbach (1988) mentioned An. ciner- eus Theobald, as An. hispaniola (Theobald), An. sergentii (Theobald), and An. superpictus larvae with Cx. mimeticus, and Azari-Hamidian et al. (2002a) collected it with An. ’hyrcanus’. An. cinereus does not exist in Iran and An. ser- gentii occurs only in southern Iran, but other species were collected with Cx. mimeticus in the current research (Table 4). Cx. pipiens larvae have been found in many different natural and artificial habitats including underground ones such as flooded cellars, drains, wells, septic tanks, underground train systems, abandoned mine tunnels, and coal mines (Hors- fall 1955, Cranston et al. 1987, Harbach 1988, Service 1993). The wide variety of the breeding places can explain why this species is one of the most abundant and common mosquito spe- cies in Iran as well as Guilan Province (Table 1 and 2). In Iran, Dow (1953), Lotfi (1970, 1976), and Yaghoobi-Ershadi et al. (1986) found this species mostly in natural habitats. Also, Zaim (1987b) collected it mainly in natural habitats (63.7%), but Mousakazemi et al. (2000) col- lected it only in rice fields (artificial habitat) and Azari-Hamidian et al. (2002a) found it mainly in man-made habitats (83.7%). In the present study, the species was collected mainly in natu- ral larval sites (67.3%), but it was collected from man-made ones (32.7%) more than any other Culex species (Table 2). It seems that the most favorable breeding places of Cx. pipiens in cities are the house ponds, wells, and the sewage wells (Golestani 1967, Lotfi 1976, Zaim 1987a). Lotfi (1976) noted the water tempera- ture and pH for Cx. pipiens 18-27 oC and 7-8 (once 6.2), respectively. Dow (1953) found Cx. Cx. territans An. claviger 26.36 An.’hyrcanus’, An. maculipennis s.l., Cx. tritaeniorhynchus 26.36 An.’hyrcanus’, An. maculipennis s.l. 15.45 Cx. pipiens 11.82 An.’hyrcanus’, An. maculipennis s.l., Cx. pipiens 9.09 An.’hyrcanus’, Cx. mimeticus, Cx. pipiens, Cx. tritaeniorhynchus 2.73 An. claviger, An. maculipennis s.l., Cx. tritaeniorhynchus 2.73 Ae. vexans, Cs. morsitans 1.82 An. claviger, An.’hyrcanus’, Ae. vexans 1.82 An. maculipennis s.l., Cx. pipiens 0.91 An. claviger, Cx. pipiens 0.91 Total 100 Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 17 pipiens larvae with An. claviger and Cx. quique- fasciatus Say, Yaghoobi-Ershadi et al. (1986) with Cx. perexiguus, as Cx. univittatus, Cx. theileri, Cx. tritaeniorhynchus, Cx. deserticola Kirkpatrick, and Cs. longiareolata, and Azari- Hamidian et al. (2002a) with An. ’hyrcanus’, An. maculipennis s.l., Ae. vexans, and Cx. tri- taeniorhynchus. Among these species, Cx. quiquefasciatus, Cx. perexiguus, and Cx. deser- ticola were not found in Guilan Province in the current study, but Cx. pipiens was with other species (Table 4). Cx. pipiens may be associ- ated with many species and share some larval sites with Cx. torrentium (Cranston et al. 1987). These species are difficult to be differentiated from each other and Cx. vagans in the larval stage (Zaim and Cranston 1986, Harbach 1988). There is little information about these two spe- cies in Iran. Cx. theileri is found in a wide range of breed- ing sites such as streams, irrigation ditches, swamps, different pools, springs, disused wells, open cisterns, and seepage water (Harbach 1988). Lotfi (1970, 1976) found it in seepages, small ponds, grassy ponds, rice fields, and agriculture water-storage pools with water temperature 16- 27 oC and pH 7-8 (once 5.4) in Iran. Dow (1953) and Yaghoobi-Ershadi et al. (1986) found it in different natural and artificial habitats. Zaim (1987b) noted that the species was mainly in natural habitats (82.1%) and Mousakazemi et al. (2000) collected it from rice fields in Isfahan Province more than any other mosquito species. Azari-Hamidian et al. (2002a) found it only in natural habitats (pools). In the present study, Cx. theileri was often collected from natural breeding sites (78.6%) and a few larvae (21.4%) were found only in rice fields as man-made ones (Table 2). The larvae of Cx. theileri were collected with An. ’hyrcanus’, An. maculipennis s.l., An. superpictus, Cx. hortensis, Cx. pipiens, Cx. tritaeniorhynchus, Cx. quiquefasciatus, Cx. perexiguus, Cx. deserticola, Cs. longiareolata, Oc. caspius s.l., and Ur. unguiculata in differ- ent areas of Iran (Dow 1953, Yaghoobi-Ershadi et al. 1986, Azari-Hamidian et al. 2002a) and Harbach (1988) mentioned that the larvae often found alone or with Cx. pipiens, Cx. perexiguus, or Cx. antennatus (Becker). In the current invest- tigation, Cx. theileri larvae were not observed alone and among these species Cx. perexiguus, Cx. quiquefasciatus, Cx. deserticola, and Cx. antennatus were not found in Guilan Province and Oc. caspius s.l. was found only in the adult stage (Azari-Hamidian et al. 2002b), but the species was associated with other species ex- cept for Ur. unguiculata (Table 4). Cx. tritaeniorhynchus is typically found in rice fields, flood waters, and marshy areas and some other natural or artificial habitats (Har- bach 1988). Lotfi (1970, 1976) found this spe- cies in rice fields, ponds, river edges, and stream edges with water temperature 19-33 oC and pH 6.4-7. Zaim (1987b) found it mostly (76%) in man-made breeding sites especially rice fields (68.5%), more than any other Culex species in Iran, but Azari-Hamidian et al.(2002a) found it mainly (83.9%) in natural habitats in Rasht County of northern Iran. This phenomenon was observed in this investigation either, and the species mainly found in natural habitats (80.6%) (Table 2). The reason probably is that there are many different natural breeding sites in Guilan Provine because of great annual rainfall, but other areas of Iran mainly have less annual rain- fall and natural breeding sites are fewer. Hors- fall (1955) mentioned many species with Cx. tritaeniorhynchus larvae. Some of them have been recorded in Iran but was not found in this study such as: An. marteri Senevet and Prun- nelle, An. sergentii, Cx. bitaeniorhynchus Giles, and Cx. vagans. Dow (1953) found Cx. tritaenio- rhynchus with An. ’hyrcanus’ and An. melanoon Hackett (as An. subalpinus Hackett and Lewis) of the Maculipennis complex, and Yaghoobi- Ershadi et al. (1986) with Cx. pipiens, Cx. theil- eri, Cx. sinaiticus Kirkpatrick, Cx. quiquefas- ciatus, Cx. perexiguus, Cx. deserticola, Oc. caspius s.l., and Ur. unguiculata in southern Iran and Azari-Hamidian et al. (2002a) with An. ’hyrcanus’, An. maculipennis s.l., Ae. vexans, and Cx. pipiens in northern Iran. Among these species Cx. sinaiticus, Cx. quiquefasciatus, Cx. perexiguus, and Cx. deserticola were not found Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 18 in this study and Oc. caspius s.l. was found only in the adult stage (Azari-Hamidian et al. 2002b). Cx. tritaeniorhynchus was associated with other species except for Ur. unguiculata (Table 4). Cx. hortensis is found in algal mats, seeps, brackish pools, domestic containers, and ce- ment vats (Horsfall 1955). Dow (1953) found it in the irrigation ditches, small, spring like pools of the river banks and shallow pools in the river beds. Zaim (1987b) found the species mostly (93.5%) in natural habitats. In the present study, this species was mainly (84.8%) found in natu- ral habitats (Table 2). Lotfi (1976) collected this species in seepages and agriculture water- storage pools with water temperature 15-26 oC and pH 6-7.5 and associated species were Cx. theileri and Cx. pipiens in northern Iran and Cx. theileri and Cx. quinquefasciatus in southern Iran. Dow (1953) found Cx. hortensis with An. maculipennis s.l., An. superpictus, Cx. theileri, and Cs. longiareolata in northwestern Iran. Zaim (1987a) stressed that the species was found mainly with An. maculipennis s.l., Cs. longiareolata, and Cs. annulata. In this study, this species was with all of these species except for Cx. quiquefasciatus that it was not found in Guilan Province (Table 4). Cx. territans is mostly found in natural swamps, ponds, and pools (Horsfall 1955). Lotfi (1973, 1976) found it in ponds with water temperature 23-27 oC and pH 7-7.5. Zaim (1987b) found it in small permanent pools. There are few records of utilization of artificial habitats including containers such as barrel for this spe- cies (Cranston et al. 1987). It is noteworthy that this species was found in man-made habitats (1.8%) such as rice fields and wells as well as natural ones (98.2%), in this study (Table 2). Cranston et al.(1987) noted this species is often in company with An. claviger. This phenome- non was exactly observed in this investigation and most of Cx. territans larvae (26.3%) were associated with this species (Table 3 and 4). Horsfall (1955) listed many species with Cx. territans larvae. Among them, An. claviger, Ae. vexans, Cx. pipiens, Cx. quinquefasciatus, and Cs. morsitans are in Iran. In the present study, Cx. quinquefasciatus was not found in Guilan Province, but other species were associated with Cx. territans (Table 4). During this investigation all reported spe- cies of the genus Culex in Guilan Provoince were collected (Zaim 1987b, Azari-Hamidian et al. 2002b), except for two records. Danilov (1975) reported Cx. torrentium from Rasht in Guilan province. The best character for identi- fication of this species from Cx. pipiens com- plex is the characters of the male hypopygia (Cranston et al. 1987, Harbach 1988). There is a unique record of Cx. quinquefasciatus in Enzeli of Guilan Province by Harbach (1988). This is out of the usual distribution of this spe- cies that occurs in central and southern Iran (Zaim 1987b). There is no more information about this record (Professor RE Harbach, per- sonal communication). The ecology of Cx. pipiens and Cx. tritaeniorhynchus, which are involved in the transmission of many pathogens to humans and domesticated animals and mak- ing nuisance, yet to be extensively studied. Lar- val habitat water temperature, pH, electric con- ductivity (EC), turbidity, and different organic and inorganic compounds are items which should be considered in forthcoming studies. All specimens of this study are deposited in the Museum of Medical Entomology at the School of Public Health, Guilan University of Medical sciences. ACKNOWLEDGEMENTS The author is grateful to Professor J Minar, Institute of Parasitology, Czech Republic, for studying specimens, Professor JF Reinert, Cen- ter for Medical, Agricultural and Veterinary Entomology, USA, for providing reprint of his articles, Professor J Jezek, Natural History Mu- seum of Prague, Czech Republic, Professor I. Gelbic, Institute of Entomology, Czech Repub- lic, Professor F Rettich, National Institute of Public Health, Czech Republic, Professor RE Harbach, the Natural History Museum, UK, for their important correspondence and assistance. Iranian J Arthropod-Borne Dis, (2007), 1(1): 9-20 S Azari-Hamidian: Larval Habitat Characteristics… 19 The author also wishes to thank Professor H Vatandoost, Department of Medical Entomol- ogy and Vector Control, School of Public Health, Tehran University of Medical Sciences, Iran, for reviewing the manuscript. This study was financially supported by Guilan University of Medical Sciences. REFERENCES Azari-Hamidian S, Yaghoobi-Ershadi MR, Javadian E (2002a) The distribution and larval habitat characteristics of mosquitoes (Diptera: Culicidae) in Rasht County (Gui- lan Province, Iran). Modarres J Med Sci. 4: 87-96 [In Persian with English abstract]. Azari-Hamidian S, Joeafshani MA, Mosslem M, Rassaei AR (2002b) Taxonomic survey of mosquitoes (Diptera: Culicidae) in Guilan Province with reporting of a subgenus new to Iranian mosquito fauna. The 15th Iranian Plant Protection Congress, 2002 September 7-11, Razi University of Kermanshah, pp. 319-320 [In Persian with English abstract]. 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